In vitro and in vivo antileishmanial activities of a Brazilian green propolis extract

The in vitro antileishmanial activity of Brazilian green propolis hydroalcoholic extract (BPE) were carried out on Leishmania (Viannia) braziliensis against both promastigote (doses ranging from 1 to 750 mu g mL(-1)) and amastigote (10, 100, and 250 mu g mL(-1)) assays in comparison with the positive (amphotericin B) and negative (dimethyl sulfoxide at 1% in physiologic solution) control groups. BPE displayed in vitro antileishmanial activities against promastigote forms of the parasite (p<0.05). However, it was inactive against its amastigote ones. In the in vitro cytotoxicity assay against Vero cells, BPE showed no cytotoxicity in the maximum doses tested. The high-performance liquid chromatography analysis allowed the identification of caffeic acid, p-coumaric acid, aromadendrine-4`-methyl-ether, 3-prenyl-p-coumaric acid (drupanin), and 3,5-diprenil-p-cumarico acid (artepillin C) as major compounds of BPE. In the in vivo assay, using a Balb/C lineage of Mus musculus male mice, groups of ten animals each were treated (1.5 mg kg day(-1)) with BPE orally (group 1), BPE topically (group 2), BPE orally and topically (group 3), and glucantime (group 4), using NaCl 0.9% (group 5) as the negative control group. Groups 1, 2, and 3 displayed a decrease on lesion development, after 90 days of treatment, by 78.6%, 84.3%, and 90.0%, respectively, while the glucantime-treated group showed 57.7% of decrease, all in comparison with the negative control group. It is the first time that the in vivo antileishmanial activity has been reported for Brazilian green propolis.

Antileishmanial, antimalarial and antimicrobial activities of the extract and isolated compounds from Austroplenckia populnea (Celastraceae)

Austroplenckia populnea (Celastraceae), known as ""marmelinho do campo"", is used in Brazilian folk medicine as antimicrobial, anti-inflammatory, and antitumoural agent. The aim of the present work was to evaluate the antimicrobial. antileishmanial and antimalarial activities of the crude hydroalcoholic extract of A. populnea (CHE) and some of its isolated compounds. The phytochemical study of the CHE was carried Out affording the isolation of methyl populnoate (1), populnoic acid (2), and stigmast-5-en-3-O-beta-(D-glucopyranoside) (3). This is the first time that the presence of compound 3 in A. populnea is reported. The results showed that the CHE presents antifungal and antibacterial activities, especially against Candida glabrata and Candida albicans, for which the CHE showed IC(50) values of 0.7 mu g mL(-1) and 5.5 mu g mL(-1), respectively, while amphotericin B showed an IC(50) value of 0.1 mu g mL(-1) against both microorganisms. Compounds 1-3 were inactive against all tested microorganisms. In the antileishmanial activity test against Leishmania donovani, the CHE showed an IC(50) value of 52 mu g mL(-1), while compounds 2 and 3 displayed an IC(50) value of 18 mu g mL(-1). In the antimalarial assay against Plasmodium falciparum (D6 and W2 clones), it was observed that all evaluated samples were inactive. In order to compare the effect on the parasites with the toxicity to mammalian cells, the cytotoxicity activity of the isolated compounds was evaluated against Vero cells, showing that all evaluated samples exhibited no cytotoxicity at the maximum dose tested.

Antimicrobial activity of the extract and isolated compounds from Baccharis dracunculifolia D. C. (Asteraceae)

Baccharis dracunculifolia D. C. (Asteraceae) is the most important plant source of the Brazilian green propolis. Since propolis is known for its antimicrobial activity, the aim of this work was to evaluate the showed that the leaves extract of B. dracunculifolia (BdE) presents antifungal and antibacterial activities, especially against Candida krusei and Cryptococcus neoformans, for which the BdE showed IC50 values of 65 mu g mL(-1) and 40 mu g mL(-1), respectively In comparison to the BdE, it was observed that the green propolis extract (GPE) showed better antimicrobial activity, displaying an IC50 value of 9 mu g mL(-1) against C krusei. Also, a phytochemical study of the BdE was carried out, affording the isolation of ursolic acid (1), 2 alpha-hydroxy-ursolic acid (2), isosakuranetin (3), aromadendrin-4`-methylether (4), baccharin (5), viscidone (6), hautriwaic acid lactone (7), and the clerodane diterpene 8. This is the first time that the presence of compounds 1, 2, and 8 in B. dracunculifolia has been reported. Among the isolated compounds, 1 and 2 showed antibacterial activity against methicillin-resistant Staphylococcus aureus, displaying IC50 values of 65 mu g mL(-1) and 40 mu g mL(-1), respectively. 3 was active against C neoformans, showing an IC50 value of 15 mu g mL(-1) and a MIC value of 40 mu g mL(-1), while compounds 4-8 were inactive against all tested microorganisms. The results showed that the BdE, similar to the GPE, displays antimicrobial activity, which may be related to the effect of several compounds present in the crude extract.

Processing of Rosmarinus officinalis Linne extract on spray and spouted bed dryers

This article presents an investigation of the potential of spray and spouted bed technology for the production of dried extracts of Rosmarinus officinalis Linne, popularly known as rosemary. The extractive solution was characterized by loss on drying, extractable matter and total phenolic and flavonoid compounds (chemical markers). The product was characterized by determination of loss on drying, size distribution, morphology, flow properties and thermal degradation and thermal behavior. The spray and spouted bed dryer performance were assessed through estimation of thermal efficiency, product accumulation and product recovery. The parameters studied were the inlet temperature of the spouting gas (80 and 150 degrees C) and the feed mass flow rate of concentrated extract relative to the evaporation capacity of the dryer, W-s/W-max (15 to 75%). The atomizing air flow rate was maintained at 20 l/min with a pressure of 196.1 kPa. The spouting gas flow rate used in the drying runs was 40% higher than the gas flow under the condition of minimum spouting. The spray drying gas flow rate was fixed at 0.0118 kg/s. Under the conditions studied, performance in the spray and spouted bed drying of rosemary extract was poor, causing high degradation of the marker compounds (mainly the phenolic compounds). Thus, process improvements are required before use on an industrial scale.

Evaluation of the genotoxic and antigenotoxic potential of Baccharis dracunculifolia extract on V79 cells by the comet assay

Baccharis dracunculifolia (Asteraceae), the main botanical source of green propolis, is a shrub of the Brazilian `cerrado`. In folk medicine it is used as an anti-inflammatory agent, mainly for the treatment of gastrointestinal diseases. The aim of the present study was to evaluate the genotoxic and antigenotoxic effects of B. dracunculifolia ethyl acetate extract (Bd-EAE) on Chinese hamster lung fibroblasts (V79 cells) by the comet assay. Methyl methanesulfonate (MMS; 200 mu M) was used as an inducer of DNA damage. Genotoxicity was evaluated using four different concentrations of Bd-EAE: 12.5, 25.0, 50.0 and 100.0 mu g ml(-1). Antigenotoxicity was assessed before, simultaneously, and after treatment with the mutagen. The results showed a significant increase in the frequency of DNA damage in cultures treated with 50.0 and 100.0 mu g ml(-1) Bd-EAE. Regarding its antigenotoxic potential, Bd-EAE reduced the frequency of DNA damage induced by MMS. However, this chemopreventive activity depended on the concentrations and treatment regimens used. The antioxidant activity of phenolic components present in Bd-EAE may contribute to reduce the alkylation damage induced by MMS. In conclusion, our findings confirmed the chemopreventive activity of Bd-EAE and showed that this effect occurs under different mechanism. Copyright (C) 2009 John Wiley & Sons, Ltd.

DRYING OF HERBAL EXTRACT IN A DRAFT-TUBE SPOUTED BED

This work evaluates the feasibility of the draft-tube spouted bed for drying of herbal extract. Drying runs were carried out according to a central composite design in a conical-cylindrical draft-tube spouted bed. The variables studied were the percentage of the drying aid (ADJ), the drying gas flow rate relative to gas flow at minimum spouting (Q/Q(ms)), and the flow rate of extract fed to the system relative to the spouting gas flow rate W(s)/W(g)). Colloidal silicon dioxide was the drying aid used in order to improve drying performance. Statistical analysis of the effects of processing parameters on product recovery, product accumulation in the bed, and product properties permitted the identification of parameters presenting significant effects on drying. Optimized drying conditions were related to experimental parameters as follow: high levels of the percentage of drying adjuvant (ADJ), high airflow rate relative to minimum spouting (Q/Q(ms)), and low values of the feed flow rate of the extract relative to the gas flow rate (W(s)/W(g)).

Anti-snake venom properties of Schizolobium parahyba (Caesalpinoideae) aqueous leaves extract

Many medicinal plants have been recommended for the treatment of snakebites. The aqueous extracts prepared from the leaves of Schizolobium parahyba (a plant found in Mata Atlantica in Southeastern Brazil) were assayed for their ability to inhibit some enzymatic and biological activities induced by Bothropspauloensis and Crotalus durissus terrificus venoms as well as by their isolated toxins neuwiedase (metalloproteinase), BnSP-7 (basic Lys49 PLA(2)) and CB (PLA(2) from crotoxin complex). Phospholipase A(2), coagulant, fibrinogenolytic, hemorrhagic and myotoxic activities induced by R pauloensis and C. d. terrificus venoms, as well as by their isolated toxins were significantly inhibited when different amounts of S. parahyba were incubated previously with these venoms and toxins before assays. However, when S. parahyba was administered at the same route as the venoms or toxins injections, the tissue local damage, such as hemorrhage and myotoxicity was only partially inhibited. The study also evaluated the inhibitory effect of S. parahyba upon the spreading of venom proteins from the injected area into the systemic circulation. The neutralization of systemic alterations induced by i.m. injection of R pauloensis venom was evaluated by measuring platelet and plasma fibrinogen levels which were significantly maintained when S. parahyba extract inoculation occurred at the same route after R pauloensis venom injection. In conclusion, the observations confirmed that the aqueous extract of S. parahyba possesses potent snake venom neutralizing properties. It may be used as an alternative treatment to serum therapy and as a rich source of potential inhibitors of toxins involved in several physiopathological human and animal diseases. Copyright (c) 2008 John Wiley & Sons, Ltd.

Neutralization of pharmacological and toxic activities of Bothrops snake venoms by Schizolobium parahyba (Fabaceae) aqueous extract and its fractions

The aqueous extract prepared from Schizolobium parahyba (Sp) leaves, a native plant from Atlantic Forest (Brazil), was tested to analyse its ability to inhibit some biological and enzymatic activities induced by Bothrops alternatus (BaltCV) and Bothrops moojeni (BmooCV) snake venoms. Sp inhibited 100% of lethality, blood incoagulability, haemorrhagic and indirect haemolytic activities at a 1:10 ratio (venom/extract, w/w), as well as coagulant activity at a 1:5 ratio (venom/extract, w/w) induced by both venoms. BaltCV fibrinogenolytic activity was also neutralized by Sp at a 1:10 ratio, resulting in total protection of fibrinogen B beta chain and partial protection of A alpha chain. Interaction tests have demonstrated that, at certain extract/proteins ratios, Sp precipitates proteins non-specifically suggesting the presence of tannins, which are very likely responsible for the excellent inhibiting effects of the analysed ophidian activities. Sp aqueous extract chromatography on Sephadex LH-20 was carried out aiming at the separation of these compounds that mask the obtained results. Thus, the fractionation of Sp resulted in three fractions: F1 (methanolic fraction); F2 (methanol:water fraction, 1:1 v/v); and F3 (aqueous fraction). These fractions were analysed for their ability to inhibit the BaltCV fibrinogenolytic activity. F1 inhibited 100% the venom fibrinogenolytic activity without presenting protein precipitation effect; F2 showed only partial inhibition of this venom activity. Finally, F3 did not inhibit fibrinogen proteolysis, but presented strong protein precipitating action. We conclude that Sp aqueous extract, together with tannins, also contains other compounds that can display specific inhibitory activity against snake venom toxins.

Use of Carnobacterium maltaromaticum cultures and hydroalcoholic extract of Lippia sidoides Cham. against Listeria monocytogenes in fish model systems

Minimally processed refrigerated ready-to-eat fishes may offer health risk of severe infection to susceptible individuals due to contamination by the psychrotolerant bacterium L monocytogenes. In this work, inhibition of L monocytogenes by a plant extract and lactic acid bacteria (IAB) was studied in model fish systems kept at 5 degrees C for 35 days. For that, fillets of tropical fish ""surubim"" (Pseudoplatystoma sp.) and hydroalcoholic extract of the plant Lippia sidoides Cham. (""alecrim pimenta"") were used. Fish peptone broth (FPB), ""surubim"" broth and ""surubim"" homogenate were inoculated with combinations of L monocytogenes and bacteriocin-producing Carnobacterium maltaromaticum (C2 and A9b(+)) and non bacteriocin-producing C. maltaromaticum (A9b(-)), in the presence or absence of extract of ""alecrim pimenta"" (EAP). In all model systems, monocultures of L monocytogenes and carnobacteria reached final populations >= 10(8) CFU/ml after 35 days, except for L monocytogenes in ""surubim"" homogenate (10(4) CFU/ml). In FPB, EAP alone and combined with cultures of LAB inhibited L monocytogenes but carnobacteria without EAP were only weakly antilisterial. In ""surubim"" broth, EAP alone did not prevent L monocytogenes growth but cultures of carnobacteria combined or not with EAP inhibited L monocytogenes, with more pronounced effect being observed for C maltaromaticum C2, which produced bacteriocin. In ""surubim"" homogenate, EAP alone and combined with cultures of C. maltaromaticum A9b(-) and A9b(+) were strongly inhibitory to L monocytogenes, while C maltaromaticum C2 with EAP caused transient inhibition of L monocytogenes. No significant inhibition of L monocytogenes was observed for carnobacteria in ""surubim"" homogenate without EAP. In conclusion, it was observed that the use of EAP and cultures of carnobacteria have potential to inhibit L monocytogenes in fish systems and the applications should be carefully studied, considering the influence of food matrix. (c) 2011 Elsevier B.V. All rights reserved.

Acute Toxicity of Schizolobium parahyba Aqueous Extract in Mice

The herbal extract of Schizolobium parahyba leaves is used commonly in the Brazil central region to treat snakebites. This study evaluates the acute toxicological effects of Schizolobium parahyba aqueous extract in mice 24 h after intraperitoneal administration. Acute toxicity was evaluated using biochemical, hematological and histopathological assays. Alterations in the levels of transaminases, bilirubin, albumin and prothrombrin time were observed, and these are likely to occur due to hepatic injury, which was confirmed by light microscopy. Liver histopathological analysis revealed the presence of lymph plasmocitary inflammatory infiltrate, but no other histopathological alterations were observed in any of the other organs analysed. The data confirm the low toxicity of the extract of Schizolobium parahyba and provide a model for the selection of a dose that does not cause injuries in the organism. Copyright (C) 2009 John Wiley & Sons, Ltd.

The antitumoral, trypanocidal and antileishmanial activities of extract and alkaloids isolated from Duguetia furfuracea

The alkaloid extract and five alkaloids isolated from subterranean stem bark of Duguetia furfuracea (Annonaceae) were investigated for the following activities: antitumoral, trypanocidal and leishmanicidal. Dicentrinone showed weak cytotoxicity, but it had the strongest leishmanicidal activity IC(50) 0.01 mu M). Duguetine and duguetine beta-N-oxide caused considerable antitumoral activity in every cell lines evaluated, although duguetine was more active against trypomastigote forms (IC(50) 9.32 mu M) than other alkaloids tested. (C) 2009 Elsevier GmbH. All rights reserved.

Evaluation of dermatological effects of cosmetic formulations containing Saccharomyces cerevisiae extract and vitamins

Saccharomyces cerevisiae extract (SCE) is used in cosmetics since it can act in oxidative stress and improve skin conditions. This study investigated dermatological effects of cosmetic formulations containing SCE and/or vitamins A, C and E. The formulation studied was supplemented or not (F1: vehicle) with vitamins A, C and E esters (F2) or with SCE (F3) or with the combination of vitamins and SCE (F4). Formulations were patch tested on back skin of volunteers. For efficacy studies, formulations were applied on volunteers and transepidermal water loss (TEWL), skin moisture (SM), skin microrelief (SMR) and free radicals protection were analysed after 3 h, 15 and 30 days of application. Volunteers were also asked about efficacy perception. It was observed that F4 provoked a slight erythema in one volunteer. All formulations enhanced forearm SM. Only F3 and F4 presented long term effects on SMR and showed higher texture values; F3 had the highest brightness values. Our results suggest that vitamins and SCE showed effects in SM and SMR. Only formulations containing SC had long term effects in the improvement of SMR. Thus, these kinds of evaluations are very important in cosmetics development to evaluate the best risk and benefit correlation. (C) 2008 Elsevier Ltd. All rights reserved.

Transformation of external sensilla to chordotonal sensilla in the cut mutant of Drosophila assessed by single-cell marking in the embryo and larva

The cut gene of Drosophila melanogaster is an identity selector gene that establishes the program of development and differentiation of external sense organs. Mutations in the cut gene cause a transformation of the external sense organs into chordotonal organs, originally assessed by the use of immunostaining methods [Bodmer et al. (1987): Cell, 51:293-307]. Because of evidence that axonal projections of the transformed neurons within the central nervous system are not completely switched in cut mutants, the transformation of the four cells making up a sense organ was reassessed using single-cell staining with fluorescent dye and differential interface contrast (DIC) microscopy of the embryo and larva. The results provide strong evidence that all cells of the sense organs are completely transformed, exhibiting the morphologies and organelles characteristic of chordotonal sense organs. A comparison of the structures of external sense organs and chordotonal organs indicates that a number of the differences could be due to the degree of development of common structures, and that cut or downstream genes modulate effector genes that are normally utilized in both receptor types. The possible derivation of insect chordotonal and external sense organs from a receptor type found in crustaceans is discussed in the light of arthropod phylogenetics and the molecular genetics of sense organ development. (C) 1997 Wiley-Liss, Inc.

A null mutation in the inflammation-associated S100 protein S100A8 causes early resorption of the mouse embryo

S100A8 (also known as CP10 or MRP8) was the first member of the S100 family of calcium-binding proteins shown to be chemotactic for myeloid cells. The gene is expressed together with its dimerization partner S100A9 during myelopoiesis in the fetal liver and in adult bone marrow as well as in mature granulocytes. In this paper we show that S100A8 mRNA is expressed without S100A9 mRNA between 6.5 and 8.5 days postcoitum within fetal cells infiltrating the deciduum in the vicinity of the ectoplacental cone. Targeted disruption of the S100A8 gene caused rapid and synchronous embryo resorption by day 9.5 of development in 100% of homozygous null embryos. Until this point there was no evidence of developmental delay in S100A8(-/-) embryos and decidualization was normal. The results of PCR genotyping around 7.5-8.5 days postcoitum suggest that the null embryos are infiltrated with maternal cells before overt signs of resorption. This work is the first evidence for nonredundant function of a member of the S100 gene family and implies a role in prevention of maternal rejection of the implanting embryo. The S100A8 null provides a new model for studying fetal-maternal interactions during implantation.

Central projections of Drosophila sensory neurons in the transition from embryo to larva

Sensory axons of different sensory modalities project into typical domains within insect ganglia. Tactile and gustatory axons project into a ventral layer of neuropil and proprioceptive afferents, including chordotonal axone, into an intermediate or dorsal layer. Here, we describe the central projections of sensory neurons in the first instar Drosophila larva, relating them to the projection of the same sensory afferents in the embryo and to sensory afferents of similar type in other insects. Several neurons show marked morphologic changes in their axon terminals in the transition between the embryo and larva. During a short morphogenetic period late in embryogenesis, the axon terminals of the dorsal bipolar dendrite stretch receptor change their shape and their distribution within the neuromere. In the larva, external sense organ neurons (es) project their axons into a ventral layer of neuropil. Chordotonal sensory neurons (ch) project into a slightly more dorsal region that is comparable to their projection in adults. The multiple dendrite (md) neurons show two distinctive classes of projection. One group of md neurons projects into the ventral-most neuropil region, the same region into which es neurons project. Members of this group are related by lineage to es neurons or share a requirement for expression of the same proneural gene during development. Other md neurons project into a more dorsal region. Sensory receptors projecting into dorsal neuropil possibly provide proprioceptive feedback from the periphery to central motorneurons and are candidates for future genetic and cellular analysis of simple neural circuitry. J. Comp. Neurol. 425:34-44, 2000. (C) 2000 Wiley-Liss, Inc.